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  Section: Biotechnology Methods » Tissue Culture Techniques
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Plant Tissue Culture demonstration by Using Somaclonal Variation to Select for Disease Resistance

Tissue Culture Techniques
  Tissue Culture Methods
  Plant Tissue Culture
  Plant Tissue Culture (Cont.)
  Many Dimensions of Plant Tissue Culture Research
  What is Plant Tissue Culture?
  Uses of Plant Tissue Culture
  Plant Tissue Culture demonstration by Using Somaclonal Variation to Select for Disease Resistance
  Demonstration of Tissue Culture for Teaching
  Preparation of Plant Tissue Culture Media
  Plant Tissue Culture Media
  Preparation of Protoplasts
  Protoplast Isolation, Culture, and Fusion
  Agrobacterium Culture and Agrobacterium — Mediated transformation
  Isolation of Chloroplasts from Spinach Leaves
  Preparation of Plant DNA using
  Suspension Culture and Production of Secondary Metabolites
  Protocols for Plant Tissue Culture
  Sterile Methods in Plant Tissue Culture
  Media for Plant Tissue Culture
  Safety in Plant Tissue Culture
  Preparation of Media for Animal Cell Culture
  Aseptic Technique
  Culture and Maintenance of Cell Lines
  Trypsinizing and Subculturing Cells from a Monolayer
  Cellular Biology Techniques
  In Vitro Methods
  Human Cell Culture Methods

Plant tissue cultures isolated from even a single cell can show variation after repeated subculture. Distinct lines can be selected with their own particular morphology and physiology. It suggests that the tissue culture contains a population of genotypes whose proportion can be altered by imposing an appropriate selection pressure. This variation can be transmitted to plants regenerated from the tissue cultures, and is called somaclonal variation. It provides an additional source of novel variation for exploitation by plant breeders.

The carrot cultivator Fancy when used in laboratory generated a series of 197 regenerant progeny lines. These
Figure 3 Scanning electron micrographs of
surface of carrot leaf 3 days after inoculation with
A. dauci.
(LHS) germination from multiseptate
conidiospore (RHS) penetration of hyphae
through the epidermal surface rather than
through stomata. Scale bar = 10 micrometers.
plants showed considerable morphological variation. They were tested for resistance to the leaf spot pathogen Alternaria dauci, which can cause total necrosis of mature leaves. They had a greater degree of variation in response than the parental cultivator, including some more resistant lines.

One symptom of the disease is loss of chlorophyl and total soluble polyphenol compounds. These reduce to a low level, 6 days after inoculation of excised leaves with A. dauci spores, when compared with uninoculated leaves. Regenerant progeny with high chlorophyl levels maintained higher chlorophyl levels after challenge with A. dauci. After self-pollinating selected high- and low-chlorophyl regenerant plants, this characteristic was inherited by their progeny, suggesting that the capacity to resist this infection is inherited.

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